Animals ; Endothelins ; physiology ; Humans ; Mice ; Pulmonary Fibrosis ; etiology ; Rats

0bjective To establish a standard curve method with more accuracy employed in fluorescence real-time PCR(RT-PCR)as a alternation of the general method.Methods β-actin and KLK11 plasmid DNA for quantitative standard curve were constructed in our study,and Plasmids of β-actin was employed as a internal control.After serial dilution these plasmid were used as DNA standard to obtained slope.Expression of these two genes in malignant prostate cancer cell line LNCAP were tested by real-time PCR,and we analyzed the RT-PCR results with two different methods and compared their accuracy.Results Thestandards curves made from these linear DNA standards showed good linearity (R2=0.991 and 0.992 for β-actin and KLK11 standards graphs),but also displayed a discrepancy in their PCR efficiency(β-actin 123% and KLK11 99%).There were different results after two different stand curve analytical method:the expression of KLK11 mRNA in LNCAP was downregulated in general standard curve method.In the new analytical method,howerer,KLK11 upregnlated for 4.46-fold.And there was a significant difference between aplification efficiency of targt gene and internal control gene(t=4.829,P<0.05).Conclusions Compared with general standard curve method,the new advanced standard curve method described here avoids an error which considers there is identical amplification efficiency between target gene and internal reference gene.It is considered to be a more correct analytical method in fluorescence real-time PCR.

OBJECTIVETo determine dopamine and its metabolites during in vivo cerebral microdialysis by routine high performance liquid chromatography with electrochemical detection.

METHODSMicrodialysis probes were placed into the right striatum of Wistar rat brains and perfused with Ringer's solution at a rate of 1.5 microL/min. A reverse phase HPLC with electrochemistry was used to assay DA, DOPAC, and HVA after cerebral microdialysates were collected every 20 minutes from awake and freely moving rats. In order to identify the reliability of this method, its selectivity, linear range, precision and accuracy were tested and the contents of DA, DOPAC, and HVA in rat microdialysates were determined.

RESULTSThe standard curve was in good linear at the concentration ranging from 74 nmol/L to 1.5 micromol/L for DOPAC (r2=0.9996), from 66 nmol/L to 1.3 micromol/L for DA (r2=1.0000) and from 69 nmol/L to 1.4 micromol/L for HVA (r2=0.9992). The recovery of DOPAC (0.30, 0.77, 1.49 micromol/L), DA (0.26, 0.69, 1.32 micromol/L), and HVA (0.27, 0.71, 1.37 micromol/L) was 82.00+/-1.70%, 104.00+/-4.00%, 98.70+/-3.10%; 92.30+/-1.50%, 105.30+/-2.30%, 108.00+/-2.00%; 80.00+/-7.80%, 107.69+/-8.00%, and 108.66+/-3.10%, respectively at each concentration. Their intra-day RSD was 3.3%, 3.4%, and 2.5%, and inter-day RSD was 4.2%, 2.3%, and 5.6%, respectively. The mean extracellular concentrations of DOPAC, DA, and HVA in rat brain microdialysates were 10.7, 2.4, and 9.2 micromol/L (n=6), respectively.

CONCLUSIONThe findings of our study suggested that the simple, accurate and stable method can be applied to basic researches of diseases related to monoamines neurotransmitters by cerebral microdialysis in rats.

Animals ; Brain ; metabolism ; Chromatography, High Pressure Liquid ; methods ; Dopamine ; metabolism ; Electrochemistry ; methods ; Microdialysis ; Rats ; Reference Standards ; Sensitivity and Specificity

Objective To analyze the clinical application of fluorescence in situ hybridization ( FISH) and karyotype analysis in prenatal diagnosis of chromosomal abnormalities .Methods The prenatal diagnosis of chromosome aneuploidies by FISH analysis of chromosome-specific probes (chromosome 13,18, 21,X,Y) in interphase amniocytes of 1 809 pregnant women of 17-38 weeks of gestational age was used , comparisons with the karyotyping results was done simultaneously.All the 1 809 cases came from Peking University First Hospital from July 1,2012 to December 31,2013, and the relevant clinical data and birth follow-up information were collected.Results All the 1 809 cases had been successfully examined by FISH , including 1 767 normal cases and 42 cases of numerical abnormality (39 cases of aneuploid, 1 case of triploid and 2 cases of mosaicism),which were consistent with the karyotyping analysis .What′s more,34 cases of chromosomal structural abnormalities , 5 cases of chimera and 12 cases of normal variant were failed to detected by FISH.Conclusion With the advantages of high-speed,simplicity,high accuracy,etc,FISH can be an effective tool in clinical applications , and had great significances in cytogenetic prenatal diagnosis combined with karyotyping analysis.

Objective To evaluate the feasibility and initial experience of laparoscopic pyelolithoto- my within renal sinus.Methods A total of 19 patients(14 men and 5 women)of nephrolithiasis on 21 sides underwent laparoscopic pyelolithotomy within renal sinus.Of them,9 cases with other upper urinary tract diseases were treated by laparoseopic pyelolithotomy at the same time.The patient age ranged from 16 to 67 years(mean,41 years).The calculi were 1.2 to 3.5 cm in diameter,and were found on the right side in 11 cases,left side in 6,and both sides in 2.Five cases concomitantly had ureteral calculi;of them,1 had bi- lateral ureter calculi with ipsilateral steinstrasse,2 had pyelic polyp(1 with giant hydronephrosis after open ureterolithotomy),and 1 had giant bydronephrosis with pyelolithiasis during pregnancy with double-J tube drainage.Twelve cases(13 sides)had experienced unsuccessful extracorporeal shock wave lithotripsy,and 1 failed with minimally invasive percutaneous nephrolithotomy.Results The procedure was successful in all the 19 cases.The operative time ranged from 75 to 240 rain(mean,115 rain).The intraoperative blood loss ranged from 30 to 100ml(mean,50 ml).One case experienced 5-d urine leakage,and was spontaneously cured.The hospital stay was 5-9 d(mean,6 d).Double-J tube drainage was 4-6 weeks.During the follow- up(3-36 months),KUB and IVU showed no upper urinary tract stricture and no residual stones except for 1 stone of 0.7 cm in the renal lower calyx in 1 case.The kidney function was improved in all cases.Con- clusions Laparoscopic pyelolithotomy is new and minimally invasive alternative approach for renal calculi. With this procedure the urinary tract complications can be treated concomitantly,thus it can partially replace open surgery.

Objective To evaluate the effects of a health education and behavior intervention project,and to provide.a scientific basis for brucellosis control.Methods By stratified cluster sampling method,target population from 6 township in Meilisi District were randomly selected to carry out baseline survey.Of these 6 townships 4 were selected as intervention townships,and two as control townships.A health education and behavior intervention program was carried out in intervention townships,and after intervention target population from 6 township were participated in the questionnaire survey.Results Awareness rate of common sense knowledge of farmers in intervention townships was 84.33％ (5237/6210) after intervention which was significantly higher than 53.56％ (2003/3740),before the intervention (x2 =112.49,P ＜ 0.01),and also higher than 54.15 ％ (1787/3300) of the control after the intervention (x2 =101.53,P ＜ 0.01).Average awareness rate(82.13％,2550/3105) of common sense on prevention after intervention was significantly higher than 58.77％(1099/1870) before the intervention (x2 =57.19,P ＜ 0.01).After intervention,average awareness rate of common sense on prevention was significantly higher than that of the average of control townships (70.36％,1161/1650,x2 =25.49,P ＜ 0.01).The awareness rate of treatment before intervention was 36.80％ (116/299),which increased to 76.36％ (436/571)after intervention (x2 =119.38,P ＜ 0.01).After the intervention the awareness rate of intervention townships was significantly higher than that of the controls (33.72％,88/261,x2 =139.69,P ＜ 0.01).In survey of these high-risk behavior in intervention townships,in addition to cattle and sheep which were regularly immunized in the intervention townships(35.91％,121/337),and the control townships(32.01％,97/303,x2 =1.08,P ＞ 0.05) and their difference was not significant,other processing such asapoblema,killing cattle,sheep,cattle to deliver,cutting the wool,regular disinfection,and accuracy of high-risk behavior protection were significantly higher than those of controls[89.83％(106/l18),91.07％(51/56),84.75％(150/177),66.67％(32/48),73.78％(242/328),51.22％(42/82),75.56％(34/45),33.69％(63/187),27.78％(15/54),21.63％(61/282),x2 value were 38.00,5.82,103.84,15.84 and 173.67,all P ＜ 0.05].Conclusion After implementation of health education and behavioral intervention,farmers brucellosis prevention knowledge and awareness of risk behaviors have significantly improved,which have achieved the desired effect.

China ; epidemiology ; Communicable Disease Control ; statistics & numerical data ; Communicable Diseases ; epidemiology ; Disease Outbreaks ; Humans

OBJECTIVETo study the absorption and accumulation of N, P and K in Ophiopgon japonicus.

METHODThe contents of N, P, K and trace element were determined respectively by using the kjeldathl method digested with H2SO4 + H2O2, ammonium vanadstemolybdate chromoscopy, flame spectrometry and atomic absorption spectrophotometry.

RESULTThe contents of N and K were the highest, and that of P was relatively lower O. japonicus. The changing pattern of N was low-high-low in the growth and development period. The accumulative speed of N had two relatively quicker and two relatively slower phases. The content of N is higher in nutritive organs; The changing pattern of P was similar to that of N, while the content of P is higher in leaves and nutritive roots at early stage of growth and development. It was high in root tuber during harvesting stage, and the accumulative quantity in the plant was increased gradually. The level of K was high in early stage and low in later stage of growth. The content of K was relatively higher in leaves and nutritive roots at early stage, and that in root tuber was slightly lower than the leaves but higher than that the nutritive roots from February to March. The accumulating quantity slightly increased in the plant.

CONCLUSIONThe application quantity of N should be increased gradually after transplant, stopped in autumn, and used again in early spring. Phosphate fertilizer is needed in autumn and winter, while large amount of potassium fertilizer shoule be used in winter tine.

Absorption ; Fertilizers ; Nitrogen ; metabolism ; Ophiopogon ; growth & development ; metabolism ; Phosphorus ; metabolism ; Plant Leaves ; metabolism ; Plant Roots ; metabolism ; Plants, Medicinal ; growth & development ; metabolism ; Potassium ; metabolism ; Seasons

OBJECTIVETo study the content of mineral elements and the principal components in Gastrodia elata.

METHODMineral elements were determined by ICP and the data was analyzed by SPSS.

RESULTK element has the highest content-and the average content was 15.31 g x kg(-1). The average content of N element was 8.99 g x kg(-1), followed by K element. The coefficient of variation of K and N was small, but the Mn was the biggest with 51.39%. The highly significant positive correlation was found among N, P and K . Three principal components were selected by principal components analysis to evaluate the quality of G. elata. P, B, N, K, Cu, Mn, Fe and Mg were the characteristic elements of G. elata.

CONCLUSIONThe content of K and N elements was higher and relatively stable. The variation of Mn content was biggest. The quality of G. elata in Guizhou and Yunnan was better from the perspective of mineral elements.

Drugs, Chinese Herbal ; analysis ; Gastrodia ; chemistry ; Minerals ; analysis ; Principal Component Analysis

OBJECTIVETo establish chromatographic fingerprint of Sichuan native medicinal plant Alisma plantago-aquatica by RP-HPLC for the quality control.

METHODThe gradient elution mode was applied in chromatographic separation, and data were analyzed by "Computer Aided Similarity Evaluation" software to compare the quality of A. plantago-aquatica samples from different habitats.

RESULTThe HPLC chromatographic fingerprinting of A. plantago-aquatica with 26 characteristic peaks was established from 17 lots of A. plantago-aquatica samples, peak 16 and 22 were identified as 24-acetyl alisol A and 23-acetyl alisol B, respectively.

CONCLUSIONThe chromatographic fingerprinting of A. plantago-aquatica with high specificity can be used to control its quality and assure lot to lot consistency. The RP-HPLC fingerprint method is repeatable, feasible in analysis of A. plantago-aquatica.

Alisma ; chemistry ; China ; Cholestenones ; analysis ; standards ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; standards ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results